冠状病毒Coronavirusl论文-1991 Identification of Coronaviruses by the Use of Indirect Protein A-Gold Immunoelectron Microscopy.pdf

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1、JVetDiagnInvest3:297-305(1991)IdentificationofcoronavirusesbytheuseofindirectproteinA-goldimmunoelectronmicroscopySergeDea,SimonGarzonAbstract.ConcentrationbyairfugeandproteinA–colloidalgoldimmunoelectronmicroscopy(PAG-IEM)offeredarapidandsensitivemethodfordetectionandidenti

2、ficationofcoronavirusesfromvariousspecies.Themethodwasappliedtopartiallypurifiedtissueculture-adaptedoregg-adaptedmammalianandaviancoronavi-rusesandtoclarifiedfecalsamplesfromdiarrheiccalvesandturkeypoultsfordetectionofentericcoronaviruses.Aggregatesofviruscoatedwithspecific

3、antibodywereseeninvirussamplesmixedwithhomologousantiserumbutnotincontrolsamplescontainingpreexposureserum.Atleasta10-50-foldenhancementofthesensitivityofdirectEMforvirusdetectionwasobtainedusingproteinA-colloidalgoldcomplexasanelectron-densemarker.ThePAG-IEMmethoddemonstrat

4、edlownonspecificbackgroundlabelingandpermitteddetectionofsolubleandparticle-associatedantigen.Reciprocalcross-reactivitywasdetectedamongthesubgroupofmam-malianhemagglutinatingcoronaviruses,andantiserato4membersofothersubgroupsonlyrecognizedtheirhomologousvirus.Coronaviruses,

5、particularlyenteropathogenicstrains,croscopytechniqueimprovesdetectionofbovineen-9fromavianandmammalianspeciesaredifficulttotericcoronavirus(BCV)infecalsamples.26Thepurposeofthepresentstudywastoevaluatethegrowincellcultures.Directelectronmicroscopy(DEM)ofviralparticlesinclar

6、ifiedfecalspecimensefficiencyofcombiningtheconcentrationofviralsus-andfluorescentantibodytechniquesfordetectionofpensionsbyairfugeandtheindirectproteinA–col-coronavirusantigensinthecellsofinfectedgutareloidalgoldimmunoelectronmicroscopylabeling(PAG-generallyusedforthediagnos

7、isofentericcoronaviralIEM)techniquesforthedetectionanddifferentiation11,14,18,19,21ofavianandmammalianentericandrespiratoryco-infections.Thedetectionofcoronavirusan-tigensbyindirectimmunofluorescence(IFF)isdepen-ronaviruses.dentontissuebeingremovedveryshortlyafterdeath,andmo

8、stantigenispresentearlyinthecourseofdis-Materialsandmethods19,21ease.Thelac